|Year : 2021 | Volume
| Issue : 3 | Page : 196-200
Periodontal manifestations as related to the cytokines in the gingival crevicular fluid
Nymphea Pandit1, Neeta Bhavsar2
1 Ph.D. Scholar, Gujarat University, Yamuna Nagar, Haryana, India
2 Department of Periodontology, Government Dental College, Ahmedabad, Gujarat, India
|Date of Submission||12-Oct-2020|
|Date of Acceptance||01-Apr-2021|
|Date of Web Publication||12-Jul-2021|
Department of Periodontology and Oral Implantology, J.N.Kapoor, D.A.V (C) Dental College, Yamuna Nagar, Haryana
Source of Support: None, Conflict of Interest: None
Backround: Periodontal diseases are classified into different categories based on the clinical symptoms and related to these there can be changes in the cytokine levels in the connective tissue and elaboration of these in the gingival crevicular fluid. (GCF). Aims and objectives: The aim of this study was to evaluate the levels of IL-11 and IL-17 in the gingival crevicular fluid of patients with Aggressive Periodontitis and Chronic Periodontitis and co-relate them with the clinical parameters. Materials and Methods: GCF was collected from the deepest sites of the periodontitis patients and ELIS A was used to determine the levels of IL-11 and IL-17 levels. Results: GCF of patients in each category was assayed and the levels of IL-11 and IL-17 were co-related with clinical symptoms. Conclusion: Study showed strong co-relation of Aggressive and Chronic Periodontitis with IL-11 and IL-17.
Keywords: Aggressive periodontitis chronic periodontitis, interleukin-11, interleukin-17
|How to cite this article:|
Pandit N, Bhavsar N. Periodontal manifestations as related to the cytokines in the gingival crevicular fluid. Indian J Dent Sci 2021;13:196-200
|How to cite this URL:|
Pandit N, Bhavsar N. Periodontal manifestations as related to the cytokines in the gingival crevicular fluid. Indian J Dent Sci [serial online] 2021 [cited 2021 Sep 20];13:196-200. Available from: http://www.ijds.in/text.asp?2021/13/3/196/321176
| Introduction|| |
Periodontitis is manifested in various forms and there have been repeated approaches to classify the diseases depending on the age of onset, severity, extent and the rate of progression. Although distinct features have been suggested to categorize the periodontitis into mainly chronic type or aggressive types, the patients do not classically fall into strict patterns. As the approach to treatment varies,, it becomes imperative to look beyond the clinical symptoms. With the advancement in understanding of the disease as well as the evaluation equipment, molecular basis of various diseases including periodontitis has been explored to confirm the diagnosis based on the clinical findings.
From the perspective of periodontitis being an immunoinflammatory disease, it makes sense to measure the level of the pro-inflammatory and anti-inflammatory mediators and compare their balance with that of the healthy patients. Among the recently explored cytokines, interleukin (IL)-11 (anti-inflammatory mediator) and IL-17 (pro-inflammatory mediator) are also involved with the regulation effect., Many studies have proven that the ratio of these two mediators can be a key factor to designate the disease to chronic type or the aggressive one. The present study was designed to evaluate the levels of the above cytokines and correlate them to the clinical features.
| Materials and Methods|| |
The proposed study is a cross-sectional study based with random sampling. A total of 100 subjects categorized into two groups of 50 subjects were selected according to the following criteria: subjects were categorized into two groups based on their diagnosis:
Group 1: –50 patients with aggressive periodontitis (age ranges from 15 to 30 years)
and Group 2: – 50 patients with chronic periodontitis (more than 40 years).
- GROUP I: For generalized aggressive periodontitis (GAgP) (n = 50), age ranges from 15 to 30 years.
- Probing depth ≥5 mm with at least 6 teeth with at least 3 of the teeth other than central incisors or first molars
- Clinical attachment level (CAL) was not consistent with the presence of local factors. Bleeding on probing was present
- Familial aggregation
- Radiographic evidence of bone loss (horizontal and vertical boss).
- GROUP II: For generalized chronic periodontitis (GCP) (n = 50), age should be <40 years.
- Patients >40 years of age
- At least 6 teeth having probing pocket depth (PPD) and CAL ≥5 mm
- Radiographic evidence of bone loss (horizontal and vertical bone loss)
- Positive bleeding on probing.
- Any use of the immunosuppressive drugs and anticonvulsant medication in the past 6 months preceding the study
- Use of antibiotics in the past 3 months or nonsteroidal anti-inflammatory drugs in the past 3 weeks.
- Any previous periodontal therapy in the past 6 months
- Pregnant or lactating female
- Patients with extensive carious lesions
- Any systemic disease (diabetes mellitus and coronary heart disease)
- Smokers and alcohol abuse.
- Plaque index (Silness and Loe, 1964)
- Sulcus bleeding index (Muhlemann and Son, 1971).
Gingival crevicular fluid (GCF) levels of IL-11 and IL-17 were evaluated as biochemical endpoints [Chart 1] and [Chart 2].
Distance exceeding 2 mm from the cement enamel junction to the alveolar crest was recorded as bone loss.
Collection of gingival crevicular fluid from the sites and processing of sample
A standardized volume of 1 μl was collected in each of two capillaries from the same site, using the calibration on white color-coded 1–5 μl calibrated volumetric capillary pipettes. The collected GCF samples were immediately transferred to airtight plastic vials (Eppendorf tubes) and stored at −70°C until assayed. After the collection of samples, samples were used for enzyme-linked immunosorbent assay (ELISA). After the collection of samples, patients were scheduled for scaling and root planning (SRP). SRP was completed in two visits within 24 h.
Patients were recalled after 2–4 weeks for follow-ups. Again the sample was collected with help of microcapillary pipette with the same procedure as described above.
The same procedure was followed for all the groups taken for the study.
Enzyme linked immunosorbent assay analysis of gingival crevicular fluid
All assay procedures were performed according to the manufacture's recommendations.
The GCF was removed from storage and placed in the kits 150 μl sample buffer (containing 1% bovine serum albumin × phosphate buffer saline) for extraction and vortexed for 1 min. The samples were incubated at + 4°C for one night and vortexed for 3 min at room temperature the following day. After that, the samples were centrifuged for 10 min at 3500 g and levels of IL-11 and IL-17 were recorded. GCF samples were analyzed for IL-17 and IL-11 using commercially available human ELISA kits. Analysis was performed according to the manufacturer's protocol. The total amount of IL-17 and IL-11 was determined in picograms (pg) and calculations of the concentration in each sample were performed by dividing the total amounts of IL-17 by the volume of the sample (pg/uml).
The study was performed on 100 subjects. Subjects were explained about the purpose and method of the study and their informed consent was obtained. Complete medical and dental history of patients was taken and clinical examination of patients was carried out. Study subjects were stratified according to clinical diagnosis. GCF samples were taken for analysis of inflammatory and immunological biomarkers. Statistical analyses were carried out to draw conclusions from periodontal biomarkers and clinical parameters between patients with generalized chronic periodontitis and generalized aggressive periodontitis.
| Results|| |
Gingival Crevicular Fluid in 50 patients was assayed and the levels of IL-11 and IL-17 showed co-relation with clinical symptoms in generalized aggressive periodontitis and generalized chronic periodontitis . other parameters like mean probing depth, CAL, plaque index and bleeding index showed non-significant difference when compared between two groups.
| Discussion|| |
The distinction between the different types of periodontitis has always been a topic of debate owing to their overlapping features in the initiation and progression of the disease. The search for the differences was then extended to the cytokines found in the milieu. There is a considerable evidence of functionally polarized responses by CD4 T helper (Th) and the CD8 T cytotoxic cell (Tc) subsets that depend on the cytokines they produce. Th1 cells produce interferon-gamma. Exciting new evidence has emerged introducing a novel subset of Th lymphocytes termed “Th17.” The role of Th17 cells and their specific cytokines (IL-17) in periodontal disease is just beginning to be investigated. Previous studies showed that IL-17 was associated with chronic periodontitis,, suggesting that it may contribute to periodontal tissue destruction.
IL-11 as an anti-inflammatory mediator has been shown to play an important role in the modulation of immune response via reduction of pro-inflammatory cytokine production in animal models., It was predicted that IL-11 may be considered as a candidate molecule for therapeutic modulation of the host response in the management of periodontal diseases., Regardless of the different clinical profiles between GAgP and GCP, it is not clear whether those patients present a distinct profile of inflammatory mediators,
The aim of this study was to determine the total amount and concentration of the cytokines IL-17 and IL-11 in the GCF of patients with GCP and GAgP. Fifty patients with GCP and 50 patients with GAgP patients were selected for the study. The power calculation of the sample size was done according to the effect size meaning thereby the difference in the two groups, the population standard deviation, the desired power of ground 95% confidence interval and the significant level. The criteria for diagnosing the patients into either of the categories were done with estimating the pocket depth, bone loss, distribution of the disease, and the age of the patient. The parameters selected for the chronic periodontitis group were 40 years or above, as the most common or prevalent age for the occurrence of this disease is 35 years and above. The average age of the patients for the aggressive periodontitis patients was 15–30 years. The present study evaluated the IL-11 and IL-17 levels in generalized chronic periodontitis patients and generalized aggressive periodontitis patients. The PPD, CAL, plaque index, and the bleeding index were evaluated as the clinical endpoints and correlated with the IL-11 and IL-17 levels. PPD represents the breakdown of the tissues around the teeth, while CAL represents the actual destruction related to the stripping of the epithelial attachment and connective tissue around the teeth. As the increasing probing depth limits the control of plaque by the patients, the plaque scores were found to be high in both the groups. The mean pocket depth in Group 1 was 6.53 ± 1.09, while it was 6.53 ± 1.09 in Group 2, and the difference was nonsignificant [Table 1]. The same was true for CAL where it was 7.13 ± 0.95 in Group 1 and 7.13 ± 0.95 in Group 2 [Table 2]. The pocket debridement leads to the decrease in the pathogen load leading to the healing and gain in the connective tissue attachment levels. The plaque index in Group I and Group II was 2.50 ± 0.50 and 2.51 ± 0.50 in Group 2 since the retention of plaque is decreased after the pocket reduction and improved ability of the patient for the plaque removal on a daily basis [Table 3]. Bleeding on probing is considered as one of the most important features that can be evaluated to assess the reduction of inflammation in the periodontal tissues., It is based on the objective evaluation rather than the subjective assessment of clinical features such as color or contour change., The bleeding index in Group I was 3.03 ± 0.58 and in Group II, it was 3.05 ± 0.58 [Table 4].
|Table 2: Comparison of mean clinical attachment level between the two groups|
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The main parameter which was evaluated between the two groups was the level of IL-11 and IL-17 in the GCF of the patients. The level of IL-11 in Group I was 52.49 ± 17.6 and in Group 2, it was 64.06 ± 26.34 [Table 5]. As IL-11 is considered to be an anti-inflammatory cytokine, it is expected to be lower in the periodontitis patients. The value was more in the chronic periodontitis patients as can be expected as compared to patients with aggressive periodontitis. IL-17 is a pro-inflammatory mediator and the levels of this cytokine are expected to be much higher in Group 1 as compared to Group 2 which was seen in our study also. The levels of IL-17 in group I were 65.80 ± 17.69 and in Group 2, the levels of IL-17 were 39.09 ± 12.17 and the difference was statistically significant [Table 6]. This type of correlation is expected as the generalized aggressive patients have an unusually severe response to the periodontal pathogens., In a study, GCF samples were collected for 30 s from two sites in 16 patients from periodontally affected sites (probing depth ≥5 mm, attachment loss ≥ mm). The comparison with healthy controls was carried out by collecting GCF samples from eight healthy volunteers and resulted in significantly lower GCF IL-17 concentration of CP patients than the healthy control group. The lower IL-17 concentration in chronic periodontitis patients than healthy controls may be depending on the higher GCF volume in diseased pockets, Gingival concentrations of both the cytokines were significantly lower in gingiva adjacent to ≥6 mm pocket. IL-11levels were significantly correlated with sulcular depth.
When the levels of IL-11 were correlated with the clinical parameters in Group 1, there was a significant positive correlation of IL-11 with the PPD and CAL [Table 7]. For Group-2, there was a significant correlation of IL-11 with PPD and CAL [Table 8]. Results have been observed by Z Yetkin et al., Johnson et al. and Ay et al. and by.
|Table 7: Correlation of interleukin 11 level with probing pocket depth and clinical attachment level in Group-1|
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|Table 8: Correlation of interleukin 11 level with probing pocket depth and clinical attachment level in Group 2|
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When the levels of IL-17 were correlated to the clinical parameters in the Group-1, there was a significant correlation of IL-17 with PPD and CAL [Table 9]. In Group-2, there was no correlation of IL-17 with PPD and CAL [Table 10].
|Table 9: Correlation of interleukin 17 level with probing pocket depth and clinical attachment level in Group 1|
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|Table 10: Correlation of interleukin 17 level with probing pocket depth and clinical attachment level in Group 2|
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| Conclusion|| |
The present study showed a statistically strong relation of the IL-11 and IL-17 with aggressive and chronic periodontitis. Higher levels of IL-11 were found to relate to healthy tissues, whereas it was vice versa for the IL-17 levels. Future studies can be undertaken to evaluate the use of IL-11 as an adjunct for the resolution of inflammation in the periodontal disease.
Ethical clearance was taken by the institutional ethical committee.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
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[Table 1], [Table 2], [Table 3], [Table 4], [Table 5], [Table 6], [Table 7], [Table 8], [Table 9], [Table 10]